关键词: 基因敲除, PARP-1, 髓母细胞瘤细胞, 神经标记物

Abstract: Objective In order to study the molecular mechanism of medulloblastoma, the cell line with PARP-1 and p53 null mutation was established and characterized. Method Mouse medulloblastoma cell line derived from PARP-1/p53 double knockout mice was established. We analyzed cell characters after 30 passages, using neuronal cell-specific markers by immunofluorescence. The cells were transfected with pEGFP-C1-Hparp-1 and pEGFP-C1 plasmids，and the expression of PARP-1 protein was detected by immunofluorescence stainning and Western blot. Result The cells showed positive immunoactivity for the neuronal-specific markers such as Vimentin, Dcx and III-Tubulin, and cells were negative for PARP-1 protein. Exogenous PARP-1 expression was visualized by immunofluorescence and Western blot analysis after pEGFP-C1-Hparp-1 transfection. Conclusions Mouse medulloblastoma cell line with defective DNA damage response has been successfully established, and provides a useful tool for further dissecting the molecular mechanism and pathogenesis of medulloblastoma.

Key words: knock-out, PARP-1gene, medulloblastoma cell, neuronal marker